IUSM-Purdue TREAT-AD Center Milestone 2 Target Enabling Package: NLRP3 (NLR family pyrin domain containing 3)

IUSM-Purdue TREAT-AD Center Milestone 2 Target Enabling Package NLRP3 (NLR family pyrin domain containing 3) Gene Symbol/Name NCBI Gene ID UniProt ID Ensembl ID NLRP3 (NLR family pyrin domain containing 3) 114548 Q96P20 ENSG00000162711 Alternatives C1orf7; CIAS1; DFNA34; AGTAVPRL; AII; AVP; FCAS; FCU; NALP3 ; PYPAF1; MWS; CLR1.1 Corresponding Author Pathum M Weerawarna Contributing Authors Omar El Jordi, Xiaoping Gu, Emily Mason, Brent Clayton, Shaoyou Chu, Timothy I . Richardson Collaborating Authors Supervision Kun Huang, Andrew Mesecar, Jeff Dage, Brent Clayton, Bruce Lamb, Alan Palkowitz, Timothy I. Richardson Date Approved by Admin Core October 15, 2025 Document version Version 1.0 Document version date October 23, 2025 Citation https://doi.org/10.5281/zenodo.17428459 Sections of this work have been and will be submitted for peer review and prospective publication Affiliations Indiana University School of Medicine, Indiana Biosciences Research Institute, Purdue University Funding U54AG065181 1. ABSTRACT The TaRget Enablement to Accelerate Therapy Development for Alzheimer’s Disease (TREAT-AD) centers were established to provide high-quality research tools and technologies to validate and advance the next generation of drug targets for Alzheimer’s disease (AD). Data, methods, and experimental resources are being openly disseminated to the AD research community to accelerate target discovery and validation. These resources are compiled as Target Enablement Packages (TEPs) and made available through the AD Knowledge Portal. Here, the IUSM-Purdue TREAT-AD Center presents a Milestone 2 (M2) TEP that evaluates the experimental feasibility of initiating a drug discovery program targeting NLRP3 by establishing and assessing initial in vitro assays and tool molecules. NLRP3 is a key sensor protein of the NLRP3 inflammasome complex that regulates caspase-1 activation and the maturation of pro-inflammatory cytokines such as IL-1β and IL-18. NLRP3 activation in microglia has been implicated in driving chronic neuroinflammation, amyloid pathology, and tau aggregation in AD. To experimentally validate literature-reported NLRP3 inhibitors and benchmark assay performance, we established paired high-content imaging assays that quantitatively measure ASC speck formation and pyroptosis in THP-1 cell models. Using these assays, we evaluated a panel of small molecules reported to directly bind the NLRP3 NACHT domain, including MCC950, Inzomelid, and two structurally distinct fused bicyclic heteroaryl chemotypes selected from a recently disclosed patent based on their potency and CNS drug-like properties. The results reproduced literature-reported activities and identified previously unreported racemic analogs of the patent selected compounds with low-nanomolar potency. Together, these findings validate our cell-based assay platform for quantitative evaluation of NLRP3 inhibitors and provide the research community with reference compounds, data, and methods to enable further investigation of NLRP3 as a therapeutic target in AD. 2. BACKGROUND NLRP3 (Nucleotide-binding domain, Leucine-rich repeat-containing family, Pyrin domain-containing 3; also known as NOD-like receptor family pyrin domain-containing 3 and officially named NLR family pyrin domain containing 3) is a protein encoded by the NLRP3 gene in humans.1 It is a member of the NOD-like receptor (NLR) family of proteins, which regulate the innate immune system. NOD-like receptors are intracellular pattern recognition receptors (PRRs) that recognize pathogen-associated molecular patterns (PAMPs), distinct from membrane-bound Toll-like receptors (TLRs) and C-type lectins (CTLs) that survey the extracellular milieu and endosomal compartments for PAMPs.2 NLRP3 is particularly known for its role in forming the NLRP3 inflammasome, a multiprotein complex that plays a critical role in inflammation and the immune response. The NLRP3 inflammasome serves as a molecular platform that activates caspase1, an enzyme responsible for the maturation and secretion of pro-inflammatory cytokines such as interleukin-1β (IL-1β) and interleukin-18 (IL-18). Activation of the NLRP3 inflammasome can be triggered by various danger signals, including PAMPs and damage-associated molecular patterns (DAMPs).3 These signals, released during cellular stress, injury, or infection, are sensed by NLRP3, leading to inflammasome assembly and subsequent immune activation.3 Figure 1. Structural organization of the NLRP3 inflammasome. (A) Schematic representation of the canonical NLRP3/AIM2 inflammasome complex, comprising a sensor molecule (NLRP3/AIM2), adaptor protein (ASC), and effector molecule (pro-caspase-1). Upon activation, NLRP3 undergoes ATP-dependent oligomerization via the NACHT domain and recruits NEK7 to form the active inflammasome complex through PYD-PYD and CARD-CARD interactions. (B) Cryo-EM structure of the NLRP3-NEK7 complex (PDB ID: 6NPY) shown in two orientations (180° rotation). Major domains including the LRR (leucine-rich repeat), NACHT (comprising HD1, WHD, NBD, and HD2), and the associated NEK7 are annotated (PYD is not included in the Cryo-EM structure). A linear domain map of NLRP3 is included at the bottom for reference. Several inflammasomes that activate caspase-1, as mentioned above, have been fully characterized and are named after the NLR or AIM2-like receptor (ALR) protein scaffolds that form their core structures, with the NLRP3 inflammasome being the most extensively studied.4 The inflammasome structure (as exemplified by NLRP3 and AIM2) generally consists of three components: a sensor molecule, an adaptor protein, and an effector molecule (Figure 1A). The sensor molecule is typically a cytosolic pattern recognition receptor (PRR), as previously described, and belongs either to the NLR family (e.g., NLRP1, NLRP3, or NLRC4) or to the ALR family (e.g., AIM2). The adaptor protein bridges the sensor and effector molecules through protein-protein interactions (PPIs). In the NLRP3 inflammasome, the adaptor protein is the apoptosis-associated speck-like protein containing a CARD (ASC), which connects the NLRP3 sensor to the pro-caspase-1 effector via PYD-PYD and CARD-CARD interactions respectively. In some inflammasomes, such as NLRP1, the adaptor protein is absent, and the sensor molecule (NLRP1) is capable of directly interacting with pro-caspase-1. The effector molecule, pro-caspase-1, interacts with ASC via its CARD domain and, upon activation and autoproteolysis, cleaves the precursor cytokines IL-1β and IL-18 into their mature forms. The detailed structure of the NLRP3 sensor molecule and the structural basis for its oligomerization, which facilitates NLRP3 inflammasome activation, have been extensively studied using cryo-EM.5 As shown in Figure 1B, the cytosolic NLRP3 receptor consists of three main domains: an Nterminal pyrin domain (PYD), a central NACHT domain, and a C-terminal leucine-rich repeat (LRR) domain. The PYD domain mediates interactions with the adaptor protein ASC via PYD-PYD interactions. The central NACHT domain comprises multiple subdomains, including the nucleotide-binding domain (NBD), helical domain 1 (HD1), winged-helix domain (WHD), and helical domain 2 (HD2). The NBD is responsible for ATP binding and hydrolysis, which is a key mediator of NLRP3 inflammasome activation. While the NLRP3 inflammasome plays a crucial role in host defense against pathogens and the maintenance of immune homeostasis, its dysregulation has been implicated in the development and progression of various inflammatory diseases, including autoinflammatory disorders, type 2 diabetes, atherosclerosis, and neurodegenerative diseases such as Alzheimer’s disease (AD), as extensively reviewed elsewhere.6 Owing to its central role in inflammation, the NLRP3 inflammasome has emerged as an important therapeutic target for the treatment of these conditions. In the early 2010s, a comprehensive review by Heneka and colleagues described the involvement of innate immune activation in the pathogenesis of neurodegenerative diseases, with a particular emphasis on AD.7 The authors summarized the literature up to 2014 and highlighted the pivotal role of NLRP3 inflammasome activation in microglia, which is triggered by misfolded host-derived proteins such as amyloid-β, leading to chronic neuroinflammation and neuronal dysfunction. In a subsequent study by the same research group, Venegas et al. provided direct experimental evidence linking NLRP3 inflammasome activation to amyloid pathology in AD.8 They demonstrated that ASC specks released by activated microglia via the NLRP3 inflammasome act as cross-seeding agents that accelerate amyloid-β aggregation and spreading in the brain. This work offers a mechanistic explanation for how innate immune activation can drive disease progression and reinforces the pathogenic role of NLRP3 inflammasome signaling in AD. Later, another study by Ising et al. demonstrated that NLRP3 inflammasome activation not only promotes amyloid pathology but also drives tau hyperphosphorylation and aggregation, thereby linking NLRP3 signaling to both hallmark pathologies of AD.9 This finding further supports the central role of microglial NLRP3 inflammasome activation in the progression of AD and highlights it as a promising therapeutic target. Drug discovery programs targeting NLRP3 were originally focused on developing broad anti-inflammatory drugs for peripheral indications. As a result, small molecule inhibitors like MCC950 generally lack central nervous system (CNS) drug-like properties. Currently there is no FDA approved drug specifically targeting NLRP3. Therefore, as an initial step, we selected from the literature a set of compounds reported to directly bind the NLRP3 NACHT domain, based on their CNS drug-like characteristics and potent inhibition of IL-1β production, and evaluated them using a pair of high-content imaging assays developed by our cellular pharmacology laboratory that quantify ASC speck formation for inflammasome activation and cellular pyroptosis, respectively. Herein, we report our findings as part of a publicly available TEP. 3. RESULTS AND DISCUSSION 3.1 NLRP3 Inflammasome Inhibition Strategies The inhibition of the NLRP3 inflammasome is best exploited by targeting its widely accepted two-step activation mechanism, which involves two sequential signals.10 Unlike other PRRs, the basal level of NLRP3 in macrophages is low, and prior to activation, its cytoplasmic expression must be upregulated, a process known as the priming step, typically initiated by Toll-like receptor (TLR) agonists such as lipopolysaccharides (LPS). This signaling leads to the activation of nuclear factor-κB (NF-κB), resulting in the upregulation of NLRP3 and pro-IL-1β expression. Therefore, one strategy for inhibiting the NLRP3 inflammasome is to target the priming step (indirect targeting) by blocking key signal transduction mediators involved in this process, such as IRAK4 and IκB kinases (IKKs), which phosphorylate the inhibitor of NF-κB (IκB) (Figure 2). Following the priming step, canonical activation of NLRP3 occurs upon recognition of an NLRP3 activator, which serves as the second signal and can range from microbial PAMPs to sterile DAMPs.6 Among these activation signals, the efflux of intracellular K⁺ was the first event identified as a key step in the NLRP3 activation process, triggered by various stimuli such as bacterial toxins, extracellular ATP, and various particulates.11 As shown in Figure 2, primed NLRP3 sensor molecules exist in an inactive, closed-cage form, as evidenced by extensive cryo-EM structural elucidation studies.5 This conformation is stabilized through PPIs mediated by LRR domains. The mitotic serine/threonine kinase NEK7 acts as a binding partner by interacting with the LRR domain of NLRP3, disrupting the oligomeric cage and stabilizing inactive NLRP3 dimers and monomers. Upon activation signaling, ATP binding and hydrolysis at the NACHT domain of NLRP3 induces a conformational change that shifts the protein into an open, active conformation. This active form subsequently undergoes inflammasome complex assembly through PPIs with other components of the inflammasome. Therefore, the second strategy for inhibiting NLRP3 inflammasome activation is to directly target the activation step, primarily through the use of small molecules that bind to the NACHT domain of NLRP3 and inhibit the ATPase activity and stabilize its closed-inactive conformation, thereby preventing the conformational change required for transition to the active-open state. Numerous small molecules that bind to the NACHT domain, such as MCC950, CY-09, tranilast, and parthenolide, have been reported in the literature, and a comprehensive review can be found elsewhere.6 In the current study, we aimed to evaluate a selected set of structurally diverse compounds that directly bind the NLRP3 NACHT domain using our in-house optimized assay, with the goal of investigating the inhibition of ASC speck formation beyond the conventional criterion of IL-1β production inhibition typically employed in NLRP3 inflammasome drug discovery campaigns. Figure 2. Therapeutic strategies for targeting the NLRP3 inflammasome. (A) Indirect inhibition (left) via blockade of the priming step through suppression of IL1R1 or TLR-mediated NF-κβ signaling (e.g. IRAK inhibition and IKKs (not shown), which reduces transcriptional upregulation of NLRP3. (B) Direct inhibition (right) of NLRP3 activation by targeting conformational dynamics. In the inactive state, NLRP3 forms closed oligomeric cage-like assemblies stabilized by LRR-LRR interactions. Activation involves NEK7 binding and ATP-dependent conformational change of the NACHT domain, leading to the open-active state. Small molecule inhibitors such as MCC950 act on the NACHT domain to stabilize the inactive conformation and inhibit the ATPase activity, preventing inflammasome assembly. 3.2 Compound Selection MCC950, one of the most widely studied NLRP3 inflammasome inhibitors, was originally discovered through a phenotype-based drug discovery campaign conducted by Pfizer in 1998, during which a diarylsulfonylurea-containing hit compound, glyburide, was found to inhibit IL-1β production in a bloodderived monocyte-based phenotypic assay.12 Subsequent optimization led to the development of MCC950, which demonstrated nanomolar potency in inhibiting IL-1β production (Figure 3).13 Figure 3. Phenotype-driven discovery and optimization of NLRP3 inflammasome inhibitor MCC950. Initial screening identified glyburide, a sulfonylurea antidiabetic, as a weak inhibitor of IL-1β production (hit). SAR efforts led to the development of diarylsulfonylurea analogs CP-412 and CP-424 with improved potency. Further chemical optimization yielded MCC950 (also known as CRID3 or CP-456), a potent and selective inhibitor of NLRP3 inflammasome activation. Photoaffinity labeling studies revealed that MCC950 directly targets the NACHT domain of NLRP3, specifically binding to the ATP hydrolysis motif (NDB) and preventing conformational activation Photoaffinity labeling experiments later confirmed that MCC950 directly binds to the NACHT domain of NLRP3.14 However, MCC950 suffers from poor brain penetration, limiting its utility to peripheral applications, and also exhibits hepatotoxicity and metabolic instability.15 These limitations prompted further optimization, ultimately yielding the MCC950-based diarylsulfonylurea clinical candidate GDC2394.16 We carefully evaluated the structure-activity relationship (SAR) studies that led to GDC-2394 in terms of IL-1β production inhibition potency and CNS drug-like properties, using a calculated CNS-MPO score. A Central Nervous System–Multiparameter Optimization (CNS-MPO) score is an empirical metric used to evaluate the drug-like properties of a molecule, combining equally weighted physicochemical parameters (calculated octanol/water partition coefficient (cLogP/cLogD), molecular weight, total polar surface area (TPSA), hydrogen-bond donors, and pKa) into a score ranging from 0 to 6. A higher score predicts that a compound is more likely to have favorable drug-like properties and CNS exposure. By balancing multiple properties rather than using rigid cutoffs based on a single parameter, the CNS-MPO score defines CNS drug-like space, guides compound selection and design and ultimately reduces attrition in the clinical development of drugs for CNS indications. We selected compounds containing the diarylsulfonylurea functionality that exhibits both potent IL-1β inhibition and a higher CNS-MPO score. As shown in Figure 4, the most potent inhibitors identified are those with IL-1β production inhibition IC50 values below 30 nM, including MCC950 and the clinical candidate GDC-2394, which exhibited lower CNS-MPO scores, suggesting limited CNS drug-like properties. However, we were able to identify a subset of compounds (compounds 3-5) with IL-1β inhibition IC50 values below 500 nM and CNS-MPO scores greater than 4. Among these, compound 5 is of particular interest, as it represents a structural isomer of Inzomelid (also known as Emlenoflast or MCC-7840; IZD174) developed by the biotech company Inflazome (acquired by Roche). Inzomelid is diarylsulfonylurea- containing NLRP3 inflammasome inhibitor with demonstrated brain exposure and potent IL-1β inhibition (<100 nM IC50).17 Based on this information, we included Inzomelid as one of the selected inhibitors that directly bind to NLRP3 with diarylsulfonylurea functionality. MCC950 was also included in this set of compounds to enable direct comparison. Figure 4. Evaluation of MCC950-based NLRP3 inhibitors. A plot of CNS-MPO score vs pIC50 values for these compounds highlights the trade-off between potency and CNS-relevant physicochemical properties. Compounds in the red-shaded region such as MCC950 and GDC-2394 (clinical candidate) are highly potent but exhibit suboptimal CNS MPO scores, whereas those in the green-shaded region (CNS-MPO > 4 and IC50 < 500 nM) display a more favorable balance. Chemical structures and IL-1β production inhibitory IC50 values are shown for representative compounds. Two compounds selected for inclusion in the current study are depicted inside gray color boxes. We also identified a set of NLRP3 inflammasome inhibitors with a novel chemotype that directly binds to the NLRP3 NACHT domain, distinct from the commonly encountered diarylsulfonylurea scaffold, exhibiting potent IL-1β production inhibitory activity, as disclosed by Aitken et al (WO 2023/066825 A1).18 These compounds are characterized by the presence of a fused bicyclic heteroaryl moiety and a phenol functional group. We performed a similar analysis to identify a subset of compounds with potent IL-1β production inhibitory activity and higher CNS-MPO scores using all the compounds that displayed <1.5 nM IC50 in the pyroptosis assay. As shown in Figure 5A, we identified two synthetically accessible compounds (example 72A and 45) with pyroptosis IC50 values below 1 nM and CNS-MPO scores greater than 3.5. Example 72A possesses the absolute stereochemistry 3aS,7aR with a cis configuration, and following the protocol described for its synthesis, we prepared compounds 6a-d corresponding to example 72A and its stereoisomers (Figure 5B). Compounds 6a and 6b represent two isomers with cis relative stereochemistry, obtained by chiral separation and denoted as cis-1 and cis-2, as the absolute stereochemistry was not assigned after chiral separation. However, as discussed later, based on the inflammasome inhibition IC50 data, we concluded that compound 6a is the analog with matching stereochemistry to example 72A. We also included the racemic version of example 72A (compound 6c) and the analog with trans relative stereochemistry (compound 6d). To evaluate the importance of the phenoxy functionality for NLRP3 inflammasome inhibitory activity, we synthesized the phenoxymethyl analogs of these compounds (compounds 7a-f), and similarly, we prepared compound 8 corresponding to example 45 along with its phenoxymethyl analog 9. We tested these compounds in a cellular assay specifically measuring inflammasome formation and pyroptosis to investigate their specific activity inhibiting inflammasome activation and a downstream pro-inflammatory form of programmed cell death. Figure 5. Identification of NLRP3 inflammasome inhibitors for CNS applications. (A) CNS-MPO versus inhibitory potency of IL-1β production (IC50) analysis of a series of NLRP3 inhibitors derived from a chemotype distinct from MCC950. The chemical structures of two compounds (examples 72A and 45) selected for the current study are shown with their absolute stereochemistry. Compounds in the greenshaded area represent candidates with an optimal balance of potency and CNS drug-like properties (CNSMPO > 3.5 and IC50 < 1 nM). (B) Compounds 6a and 6b represent two isomers with cis relative stereochemistry, obtained by chiral separation and denoted as cis-1 and cis-2, as the absolute stereochemistry was not assigned in this experiment. Based on the inflammasome inhibition data (IC50), we concluded that compound 6a (highlighted in blue) is the analog with matching stereochemistry to example 72A. Compounds 6c and 6d are the racemic versions of example 72A with cis and trans relative stereochemistry, respectively. Compounds 7a-7f represent the phenoxymethylated versions. Compound 8 corresponds to example 45, and its phenoxymethylated analog is compound 9. (5) Yu, X.; Matico, R. E.; Miller, R.; Chauhan, D.; Van Schoubroeck, B.; Grauwen, K.; Suarez, J.; Pietrak, B.; Haloi, N.; Yin, Y.; et al. Structural basis for the oligomerization-facilitated NLRP3 activation. Nat Commun 2024, 15 (1), 1164. DOI: 10.1038/s41467-024-45396-8 From NLM Medline. (6) Ma, Q. Pharmacological Inhibition of the NLRP3 Inflammasome: Structure, Molecular Activation, and Inhibitor-NLRP3 Interaction. Pharmacol Rev 2023, 75 (3), 487-520. DOI: 10.1124/pharmrev.122.000629 From NLM Medline. (7) Heneka, M. T.; Kummer, M. P.; Latz, E. Innate immune activation in neurodegenerative disease. Nat Rev Immunol 2014, 14 (7), 463-477. DOI: 10.1038/nri3705 From NLM Medline. (8) Venegas, C.; Kumar, S.; Franklin, B. S.; Dierkes, T.; Brinkschulte, R.; Tejera, D.; Vieira-Saecker, A.; Schwartz, S.; Santarelli, F.; Kummer, M. P.; et al. Microglia-derived ASC specks cross-seed amyloid-beta in Alzheimer's disease. Nature 2017, 552 (7685), 355-361. DOI: 10.1038/nature25158 From NLM Medline. (9) Ising, C.; Venegas, C.; Zhang, S.; Scheiblich, H.; Schmidt, S. V.; Vieira-Saecker, A.; Schwartz, S.; Albasset, S.; McManus, R. M.; Tejera, D.; et al. NLRP3 inflammasome activation drives tau pathology. Nature 2019, 575 (7784), 669-673. DOI: 10.1038/s41586-019-1769-z From NLM Medline. (10) McKee, C. M.; Coll, R. C. NLRP3 inflammasome priming: A riddle wrapped in a mystery inside an enigma. J Leukoc Biol 2020, 108 (3), 937-952. DOI: 10.1002/JLB.3MR0720-513R From NLM Medline. (11) Swanson, K. V.; Deng, M.; Ting, J. P. The NLRP3 inflammasome: molecular activation and regulation to therapeutics. Nat Rev Immunol 2019, 19 (8), 477-489. DOI: 10.1038/s41577-019-0165-0 From NLM Medline. (12) Perregaux, D. G.; McNiff, P.; Laliberte, R.; Hawryluk, N.; Peurano, H.; Stam, E.; Eggler, J.; Griffiths, R.; Dombroski, M. A.; Gabel, C. A. Identification and characterization of a novel class of interleukin-1 posttranslational processing inhibitors. J Pharmacol Exp Ther 2001, 299 (1), 187-197. From NLM Medline. (13) Coll, R. C.; Robertson, A. A.; Chae, J. J.; Higgins, S. C.; Munoz-Planillo, R.; Inserra, M. C.; Vetter, I.; Dungan, L. S.; Monks, B. G.; Stutz, A.; et al. A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases. Nat Med 2015, 21 (3), 248-255. DOI: 10.1038/nm.3806 From NLM Medline. (14) Coll, R. C.; Hill, J. R.; Day, C. J.; Zamoshnikova, A.; Boucher, D.; Massey, N. L.; Chitty, J. L.; Fraser, J. A.; Jennings, M. P.; Robertson, A. A. B.; et al. MCC950 directly targets the NLRP3 ATP-hydrolysis motif for inflammasome inhibition. Nat Chem Biol 2019, 15 (6), 556-559. DOI: 10.1038/s41589-019-0277-7 From NLM Medline. (15) Hill, J. R.; Shao, X.; Massey, N. L.; Stauff, J.; Sherman, P. S.; Robertson, A. A. B.; Scott, P. J. H. Synthesis and evaluation of NLRP3-inhibitory sulfonylurea [(11)C]MCC950 in healthy animals. Bioorg Med Chem Lett 2020, 30 (12), 127186. DOI: 10.1016/j.bmcl.2020.127186 From NLM Medline. (16) McBride, C.; Trzoss, L.; Povero, D.; Lazic, M.; Ambrus-Aikelin, G.; Santini, A.; Pranadinata, R.; Bain, G.; Stansfield, R.; Stafford, J. A.; et al. Overcoming Preclinical Safety Obstacles to Discover (S)-N-((1,2,3,5,6,7Hexahydro-s-indacen-4-yl)carbamoyl)-6-(methylamino)-6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine-3sulfonamide (GDC-2394): A Potent and Selective NLRP3 Inhibitor. J Med Chem 2022, 65 (21), 14721-14739. DOI: 10.1021/acs.jmedchem.2c01250 From NLM Medline. (17) Miller, D.; Macleod, A.; Del, R.; Stratford, S. Preparation of a sodium salt of N-((1, 2, 3, 5, 6, 7hexahydro-s-indacen-4-yl) carbamoyl)-1-isopropyl-1H-pyrazole-3-sulfonamide WO2019206871. Inflazome Limited 2019. (18) AITKEN, L. S.; BOUCHE, L. A.; Guba, W.; Jaeschke, G.; JOHNSTON, H. J.; MESCH, S. K.; Patiny-Adam, A.; Schnider, C.; SHANNON, J. M.; Steiner, S. Fused bicyclic heteroaryl compounds useful as nlrp3 inhibitors. WO 2023/066825 A1. (19) Vande Walle, L.; Lamkanfi, M. Drugging the NLRP3 inflammasome: from signalling mechanisms to therapeutic targets. Nat Rev Drug Discov 2024, 23 (1), 43-66. DOI: 10.1038/s41573-023-00822-2 From NLM Medline. (20) Nizami, S.; Millar, V.; Arunasalam, K.; Zarganes-Tzitzikas, T.; Brough, D.; Tresadern, G.; Brennan, P. E.; Davis, J. B.; Ebner, D.; Di Daniel, E. A phenotypic high-content, high-throughput screen identifies inhibitors of NLRP3 inflammasome activation. Sci Rep 2021, 11 (1), 15319. DOI: 10.1038/s41598-021-94850-w From NLM Medline. (21) Stanton, C.; Sun, J.; Nutsch, K.; Rosarda, J. D.; Nguyen, T.; Li-Ma, C.; Njomen, E.; Melillo, B.; Kutseikin, S.; Saez, E.; et al. Covalent Targeting As a Common Mechanism for Inhibiting NLRP3 Inflammasome Assembly. ACS Chem Biol 2024, 19 (2), 254-265. DOI: 10.1021/acschembio.3c00330 From NLM Medline.